Aryl and heterocyclyl substituted pyrimidine derivatives as anti-coagulants

ABSTRACT

This invention is directed to aryl and heterocyclyl substituted pyrimidine derivatives selected from the following formulae: wherein Z1, Z2, R1, R2, R3, R4, R5 and R6 are defined herein. These compounds are useful as anti-coagulants.

FIELD OF THE INVENTION

The present invention is directed to aryl and heterocyclyl substitutedpyrimidine derivatives and their pharmaceutically acceptable salts,which inhibit the enzyme, factor Xa, thereby being useful asanti-coagulants. It also relates to pharmaceutical compositionscontaining the derivatives or their pharmaceutically acceptable salts,and methods of their use.

BACKGROUND OF THE INVENTION

Factor Xa is a member of the trypsin-like serine protease class ofenzymes. A one-to-one binding of factors Xa and Va with calcium ions andphospholipid forms the prothrombinase complex which converts prothrombinto thrombin. Thrombin, in turn, converts fibrinogen to fibrin whichpolymerizes to form insoluble fibrin.

In the coagulation cascade, the prothrombinase complex is the convergentpoint of the intrinsic (surface activated) and extrinsic (vesselinjury-tissue factor) pathways (Biochemistry (1991), Vol. 30, p. 10363;and Cell (1988), Vol. 53, pp. 505-518). The model of the coagulationcascade has been refined further with the discovery of the mode ofaction of tissue factor pathway inhibitor (TFPI) (Seminars in Hematology(1992), Vol. 29, pp. 159-161). TFPI is a circulating multi-domain serineprotease inhibitor with three Kunitz-type domains which competes withfactor Va for free factor Xa. Once formed, the binary complex of factorXa and TFPI becomes a potent inhibitor of the factor VIIa and tissuefactor complex.

Factor Xa can be activated by two distinct complexes, by tissuefactor-VIIa complex on the "Xa burst" pathway and by the factorIXa-VIIIa complex (TENase) of the "sustained Xa" pathway in thecoagulation cascade. After vessel injury, the "Xa burst" pathway isactivated via tissue factor (TF). Up regulation of the coagulationcascade occurs via increased factor Xa production via the "sustained Xa"pathway. Down regulation of the coagulation cascade occurs with theformation of the factor Xa-TFPI complex, which not only removes factorXa but also inhibits further factor formation via the "Xa burst"pathway. Therefore, the coagulation cascade is naturally regulated byfactor Xa.

The primary advantage of inhibiting factor Xa over thrombin in order toprevent coagulation is the focal role of factor Xa versus the multiplefunctions of thrombin. Thrombin not only catalyzes the conversion offibrinogen to fibrin, factor VIII to VIIIA, factor V to Va, and factorXI to XIa, but also activates platelets, is a monocyte chemotacticfactor, and mitogen for lymphocytes and smooth muscle cells. Thrombinactivates protein C, the in vivo anti-coagulant inactivator of factorsVa and VIIIa, when bound to thrombomodulin. In circulation, thrombin israpidly inactivated by antithrombin III (ATIII) and heparin cofactor II(HCII) in a reaction which is catalyzed by heparin or otherproteoglycan-associated glycosaminoglycans, whereas thrombin in tissuesis inactivated by the protease, nexin. Thrombin carries out its multiplecellular activation functions through a unique "tethered ligand"thrombin receptor (Cell (1991), Vol. 64, p. 1057), which requires thesame anionic binding site and active site used in fibrinogen binding andcleavage and by thrombomodulin binding and protein C activation. Thus, adiverse group of in vivo molecular targets compete to bind thrombin andthe subsequent proteolytic events will have very different physiologicalconsequences depending upon which cell type and which receptor,modulator, substrate or inhibitor binds thrombin.

Published data with the proteins antistasin and tick anti-coagulantpeptide (TAP) demonstrate that factor Xa inhibitors are efficaciousanti-coagulants (Thrombosis and Haemostasis (1992), Vol. 67, pp.371-376; and Science (1990), Vol. 248, pp. 593-596).

The active site of factor Xa can be blocked by either a mechanism-basedor a tight binding inhibitor (a tight binding inhibitor differs from amechanism-based inhibitor by the lack of a covalent link between theenzyme and the inhibitor). Two types of mechanism-based inhibitors areknown, reversible and irreversible, which are distinguished by ease ofhydrolysis of the enzyme-inhibitor link (Thrombosis Res. (1992), Vol.67, pp. 221-231; and Trends Pharmacol. Sci. (1987), Vol. 8, pp.303-307). A series of guanidino compounds are examples of tight-bindinginhibitors (Thrombosis Res. (1980), Vol. 19, pp. 339-349).Arylsulfonyl-arginine-piperidine-carboxylic acid derivatives have alsobeen shown to be tight-binding inhibitors of thrombin (Biochem. (1984),Vol. 23, pp. 85-90), as well as a series of arylamidine-containingcompounds, including 3-amidinophenylaryl derivatives (Thrombosis Res.(1983), Vol. 29, pp. 635-642) and bis(amidino)benzyl cycloketones(Thrombosis Res. (1980), Vol. 17, pp. 545-548). However, these compoundsdemonstrate poor selectivity for factor Xa.

RELATED DISCLOSURES

European Published Patent Application 0 540 051 (Nagahara et al.)describes aromatic amidine derivatives. These derivatives are stated tobe capable of showing a strong anticoagulant effect through reversibleinhibition of factor Xa.

The synthesis of α,α'-bis(amidinobenzylidene)cycloalkanones andα,α'-bis(amidinobenzyl)cycloalkanones is described in Pharmazie (1977),Vol. 32, No. 3, pp. 141-145. These compounds are disclosed as beingserine protease inhibitors.

U.S. Pat. No. 5,451,700 (Morrissey et al.) describes amidino compounds.These compounds are stated to be useful as selective LTB₄ receptorantagonists.

U.S. Pat. No. 5,612,363 (Mohan et al.) describes N,N-di(aryl) cyclicurea derivatives. These compounds are stated to be factor Xa inhibitors,thereby being useful as anticoagulants.

U.S. Pat. No. 5,633,381 (Dallas et al.) describes (Z,Z), (Z,E) and (E,Z)isomers of substituted bis(phenylmethylene)cycloketones. These compoundsare disclosed as being factor Xa inhibitors, thereby being useful asanticoagulants.

PCT Published Patent Application WO/96/28427 (Buckman et al.) describesbenzamidine derivatives. These compounds are stated to be factor Xainhibitors, thereby being useful as anticoagulants.

PCT Published Patent Application WO/97/21437 (Arnaiz et al.) describesnaphthyl-substituted benzimidazole derivatives. These compounds aredisclosed as being factor Xa inhibitors, thereby being useful asanticoagulants.

PCT Published Patent Application WO/97/29067 (Kochanny et al.) describesbenzamidine derivatives that are substituted by amino acid and hydroxyacid derivatives. These compounds are stated to be factor Xa inhibitors,thereby being useful as anticoagulants.

The above references, published patent applications and U.S. patents areherein incorporated in full by reference.

SUMMARY OF THE INVENTION

This invention is directed to compounds or their pharmaceuticallyacceptable salts which inhibit human factor Xa and are therefore usefulas pharmacological agents for the treatment of disease-statescharacterized by thrombotic activity.

Accordingly, in one aspect, this invention provides compounds selectedfrom the group consisting of the following formulae: ##STR2## wherein:Z¹ is --O--, --N(R⁷)--, --CH₂ O-- or --S(O)_(n) -- (where n is 0 to 2);

Z² is --O--, --N(R⁷)--, --OCH₂ -- or --S(O)_(n) -- (where n is 0 to 2);

R¹ and R⁴ are each independently hydrogen, halo, alkyl, nitro, --OR⁷,--C(O)OR⁷, --C(O)N(R⁷)R⁸, --N(R⁷)R⁸, --N(R⁷)C(O)R⁷, or --N(H)S(O)₂ R⁹ ;

R² is --C(NH)NH₂, --C(NH)N(H)OR⁷, --C(NH)N(H)C(O)OR⁹, --C(NH)N(H)C(O)R⁷,--C(NH)N(H)S(O)₂ R⁹, or --C(NH)N(H)C(O)N(H)R⁷ ;

R³ is hydrogen, halo, alkyl, haloalkyl, nitro, ureido, guanidino, --OR⁷,--C(NH)NH₂, --C(NH)N(H)OR⁷, --C(O)N(R⁷)R⁸, --R¹⁰ --C(O)N(R⁷)R⁸,--CH(OH)C(O)N(R⁷)R⁸, --N(R⁷)R⁸, --R¹⁰ --N(R⁷)R⁸, --C(O)OR⁷, --R¹⁰--C(O)OR⁷, --N(R⁷)C(O)R⁷, (1,2)-tetrahydropyrimidinyl (optionallysubstituted by alkyl), (1,2)-imidazolyl (optionally substituted byalkyl), or (1,2)-imidazolinyl (optionally substituted by alkyl);

each R⁵ are independently hydrogen, halo, alkyl, haloalkyl, nitro,aralkoxy, --OR⁹, --R¹⁰ --OR⁹, --N(R⁷)R⁸, --C(O)O⁷, --R¹⁰ --C(O)OR⁷,--C(O)N(R⁷)R⁸, --R¹⁰ --C(O)N(R⁷)R⁸, --C(O)N(R⁷)CH₂ C(O)N(R⁷)R⁸,--N(R⁷)C(O)N(R⁷)R⁸, --N(R⁷)C(O)R⁸, --N(R⁷)S(O)₂ R⁹, or--N(R⁷)C(O)N(R⁷)--CH₂ C(O)N(R⁷)R⁸ ;

each R⁶ is aryl (optionally substituted by halo, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), aralkyl (wherein the aryl is optionallysubstituted by halo, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy,amino, dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl),heterocyclyl (optionally substituted by halo, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), or heterocyclylalkyl (wherein the heterocyclyl isoptionally substituted by halo, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl);

each R⁷ and R⁸ is independently hydrogen, alkyl, aryl (optionallysubstituted by halo, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy,amino, dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), oraralkyl (wherein the aryl is optionally substituted by halo, alkyl,aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino, dialkylamino,monoalkylamino, nitro, carboxy, alkoxycarbonyl, aminocarbonyl,monoalkylaminocarbonyl, or dialkylaminocarbonyl);

each R⁹ is alkyl, aryl (optionally substituted by halo, alkyl, aryl,aralkyl, hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino,nitro, carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl,or dialkylaminocarbonyl), or aralkyl (wherein the aryl is optionallysubstituted by halo, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy,amino, dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), and

each R¹⁰ is independently an alkylene or alkylidene chain;

as a single stereoisomer or a mixture thereof; or a pharmaceuticallyacceptable salt thereof.

In another aspect, this invention provides compositions useful intreating a human having a disease-state characterized by thromboticactivity, which composition comprises a therapeutically effective amountof a compound of the invention as described above, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable excipient.

In another aspect, this invention provides a method of treating a humanhaving a disease-state characterized by thrombotic activity, whichmethod comprises administering to a human in need thereof atherapeutically effective amount of a compound of the invention asdescribed above.

In another aspect, this invention provides a method of treating a humanhaving a disease-state alleviated by the inhibition of factor Xa, whichmethod comprises administering to a human in need thereof atherapeutically effective amount of a compound of the invention asdescribed above.

In another aspect, this invention provides a method of inhibiting humanfactor Xa in vitro by the administration of a compound of the invention.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated:

"Alkyl" refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to six carbon atoms, and which is attachedto the rest of the molecule by a single bond, e.g., methyl, ethyl,n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl,1,1-dimethylethyl (t-butyl), and the like.

"Alkoxy" refers to a radical of the formula --OR_(a) where R_(a) is analkyl radical as defined above, e.g., methoxy, ethoxy, n-propoxy,1-methylethoxy (iso-propoxy), n-butoxy, n-pentoxy, 1,1-dimethylethoxy(t-butoxy), and the like.

"Alkoxycarbonyl" refers to a radical of the formula --C(O)OR_(a) whereR_(a) is an alkyl radical as defined above, e.g., methoxycarbonyl,ethoxycarbonyl, n-propoxycarbonyl, 1-methylethoxycarbonyl(isopropoxycarbonyl), n-butoxycarbonyl, n-pentoxycarbonyl,1,1-dimethylethoxycarbonyl (t-butoxycarbonyl), and the like.

"Alkanol" refers to a compound of the formula R_(a) --OH where R_(a) isan alkyl radical as defined above, e.g., methanol, ethanol, n-propanol,etc.

"Alkylene chain" refers to straight or branched chain divalent radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation and having from one to six carbon atoms, e.g., methylene,ethylene, propylene, n-butylene and the like.

"Alkylidene chain" refers to a straight or branched chain unsaturateddivalent radical consisting solely of carbon and hydrogen atoms, havingfrom one to six carbon atoms, wherein the unsaturation is present onlyas double bonds and wherein a double bond can exist between the firstcarbon of the chain and the rest of the molecule, e.g., ethylidene,propylidene, n-butylidene, and the like.

"Aryl" refers to a phenyl or naphthyl radical.

"Aralkyl" refers to a radical of the formula --R_(a) R_(b) where R_(a)is an alkyl radical as defined above, substituted by R_(b), an arylradical, as defined above, e.g., benzyl.

"Aralkoxy" refers to a radical of the formula --OR_(c) where R_(c) is anaralkyl radical as defined above, e.g., benzyloxy, and the like.

"Amidino" refers to the radical --C(NH)NH₂.

"Aminocarbonyl" refers to the radical --C(O)NH₂.

"Dialkylamino" refers to a radical of the formula --N(R_(a))R_(a) whereeach R_(a) is independently an alkyl radical as defined above, e.g.,dimethylamino, methylethylamino, diethylamino, dipropylamino,ethylpropylamino, and the like.

"Dialkylaminocarbonyl" refers to a radical of the formula--C(O)N(R_(a))R_(a) where each R_(a) is independently an alkyl radicalas defined above, e.g., dimethylaminocarbonyl, methylethylaminocarbonyl,diethylaminocarbonyl, dipropylaminocarbonyl, ethylpropylaminocarbonyl,and the like.

"Halo" refers to bromo, chloro, iodo or fluoro.

"Haloalkyl" refers to an alkyl radical, as defined above, that issubstituted by one or more halo radicals, as defined above, e.g.,trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl,1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl,1-bromomethyl-2-bromoethyl, and the like.

"Heterocyclyl" refers to a stable 3- to 15-membered ring radical whichconsists of carbon atoms and from one to five heteroatoms selected fromthe group consisting of nitrogen, phosphorus, oxygen and sulfur. Forpurposes of this invention, the heterocyclyl radical is not attached topyrimidinyl moiety of the compounds of formula (I), (II) and (III)through a hetero atom. In addition, for purposes of this invention, theheterocyclyl radical may be a monocyclic, bicyclic or tricyclic ringsystem, which may include fused or bridged ring systems, and thenitrogen, phosphorus, carbon or sulfur atoms in the heterocyclyl radicalmay be optionally oxidized to various oxidation states. In addition, thenitrogen atom in the heterocyclyl radical may be optionally quaternized;and the heterocyclyl radical may be partially or fully saturated oraromatic. Examples of such heterocyclyl radicals include, but are notlimited to, azetidinyl, acridinyl, benzodioxolyl, benzodioxanyl,benzofuranyl, carbazoyl, cinnolinyl, dioxolanyl, indolizinyl,naphthyridinyl, perhydroazepinyl, phenazinyl, phenothiazinyl,phenoxazinyl, phthalazinyl, pteridinyl, purinyl, quinazolinyl,quinoxalinyl, quinolinyl, isoquinolinyl, tetrazoyl,tetrahydroisoquinolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, pyrrolyl,4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl,imidazolinyl, imidazolidinyl, dihydropyridinyl, tetrahydropyridinyl.pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolinyl,oxazolidinyl, triazolyl, indanyl, isoxazolyl, isoxazolidinyl,morpholinyl, thiazolyl, thiazolinyl, thiazolidinyl, isothiazolyl,quinuclidinyl, isothiazolidinyl, indolyl, isoindolyl, indolinyl,isoindolinyl, octahydroindolyl, octahydroisoindolyl, quinolyl,isoquinolyl, decahydroisoquinolyl, benzimidazolyl, thiadiazolyl,benzopyranyl, benzothiazolyl, benzoxazolyl, furyl, tetrahydrofuryl,tetrahydropyranyl, thienyl, benzothienyl, thiamorpholinyl,thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, dioxaphospholanyland oxadiazolyl.

"Heterocyclylalkyl" refers to a radical of the formula --R_(a) --R_(d)where R_(a) is an alkyl radical as defined above and R_(d) is aheterocyclyl radical as defined above, for example,(4-methylpiperazin-1-yl)methyl, (morpholin-4-yl)methyl,2-(oxazolin-2-yl)ethyl, and the like.

"(1,2)-Imidazolyl" refers to an imidazolyl radical attached at eitherthe 1- or 2-position.

"(1,2)-Imidazolinyl" refers to a 4,5-dihydroimidazolyl radical attachedat either the 1- or the 2-position.

"Monoalkylamino" refers to a radical of the formula --NHR_(a) whereR_(a) is an alkyl radical as defined above, e.g., methylamino,ethylamino, propylamino, and the like.

"Monoalkylaminocarbonyl" refers to a radical of the formula--C(O)NHR_(a) where R_(a) is an alkyl radical as defined above, e.g.,methylaminocarbonyl, ethylaminocarbonyl, propylaminocarbonyl, and thelike.

"(1,2)-Tetrahydropyrimidinyl" refers to a tetrahydropyrimidinyl radicalattached at either the 1- or 2-position.

"Optional" or "optionally" means that the subsequently described eventof circumstances may or may not occur, and that the description includesinstances where said event or circumstance occurs and instances in whichit does not. For example, "optionally substituted aryl" means that thearyl radical may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution.

"Pharmaceutically acceptable salt" includes both acid and base additionsalts.

"Pharmaceutically acceptable acid addition salt" refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid and the like, andorganic acids such as acetic acid, trifluoroacetic acid, propionic acid,glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid,succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid,cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like.

"Pharmaceutically acceptable base addition salt" refers to those saltswhich retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Salts derived from inorganic bases include, but are notlimited to, the sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Preferred inorganic salts are the ammonium, sodium, potassium, calcium,and magnesium salts. Salts derived from organic bases include, but arenot limited to, salts of primary, secondary, and tertiary amines,substituted amines including naturally occurring substituted amines,cyclic amines and basic ion exchange resins, such as isopropylamine,trimethylamine, diethylamine, triethylamine, tripropylamine,ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperazine, piperidine,N-ethylpiperidine, polyamine resins and the like. Particularly preferredorganic bases are isopropylamine, diethylamine, ethanolamine,trimethylamine, dicyclohexylamine, choline and caffeine.

"Therapeutically effective amount" refers to that amount of a compoundof the invention which, when administered to a human in need thereof, issufficient to effect treatment, as defined below, for disease-statescharacterized by thrombotic activity. The amount of a compound of theinvention which constitutes a "therapeutically effective amount" willvary depending on the compound, the disease-state and its severity, andthe age of the human to be treated, but can be determined routinely byone of ordinary skill in the art having regard to his own knowledge andto this disclosure.

"Treating" or "treatment" as used herein covers the treatment of adisease-state in a human, which disease-state is characterized bythrombotic activity, and includes:

(i) preventing the disease-state from occurring in a human, inparticular, when such human is predisposed to the disease-state but hasnot yet been diagnosed as having it;

(ii) inhibiting the disease-state, i.e., arresting its development; or

(iii) relieving the disease-state, i.e., causing regression of thedisease-state.

The yield of each of the reactions described herein is expressed as apercentage of the theoretical yield.

The compounds of the invention, or their pharmaceutically acceptablesalts, may have asymmetric carbon atoms, oxidized sulfur atoms orquaternized nitrogen atoms in their structure. The compounds of theinvention and their pharmaceutically acceptable salts may thereforeexist as single stereoisomers, racemates, and as mixtures of enantiomersand diastereomers. The compounds may also exist as geometric isomers.All such single stereoisomers, racemates and mixtures thereof, andgeometric isomers are intended to be within the scope of this invention.

The nomenclature used herein is a modified form of the I.U.P.A.C. systemwherein the compounds of the invention are named as pyrimidinylderivatives of benzamidine. For example, a compound of the inventionselected from formula (I): ##STR3## where Z¹ and Z² are both --O--, R¹is hydroxy, R² is amidino, R³ is dimethylaminocarbonyl, R⁴ is hydrogen,R⁵ is hydrogen, and R⁶ is 4-methylphenyl is named herein as4-hydroxy-3-[[6-(3-(dimethylaminocarbonyl)phenoxy)-2-(4-methylphenyl)pyrimidin-4-yl]oxy]benzamidine.

For purposes of this invention, parenthesis are used to denotesubstituents of a main atom of the substituent. For example,--C(NH)N(H)C(O)OR⁹ refers to the substituent of the formula: ##STR4##Utility and Administration

A. Utility

The compounds of the invention are inhibitors of the serine protease,factor Xa, and are therefore useful in disease-states characterized bythrombotic activity based on factor Xa's role in the coagulation cascade(see Background of the Invention above). A primary indication for thecompounds is prophylaxis for long term risk following myocardialinfarction. Additional indications are prophylaxis of deep veinthrombosis (DVT) following orthopedic surgery or prophylaxis of selectedpatients following a transient ischemic attack. The compounds of theinvention may also be useful for indications in which coumadin iscurrently used, such as for DVT or other types of surgical interventionsuch as coronary artery bypass graft and percutaneous transluminalcoronary angioplasty. The compounds are also useful for the treatment ofthrombotic complications associated with acute promyelocytic leukemia,diabetes, multiple myelomas, disseminated intravascular coagulationassociated with septic shock, purpura fulminanas associated infection,adult respiratory distress syndrome, unstable angina, and thromboticcomplications associated with aortic valve or vascular prosthesis. Thecompounds are also useful for prophylaxis for thrombotic diseases, inparticular in patients who have a high risk of developing such disease.

In addition, the compounds of the invention are useful as in vitro andin vivo diagnostic reagents for selectively inhibiting factor Xa withoutinhibiting other components of the coagulation cascade.

B. Testing

The primary bioassays used to demonstrate the inhibitory effect of thecompounds of the invention on factor Xa are simple chromogenic assaysinvolving only serine protease, the compound of the invention to betested, substrate and buffer (see, e.g., Thrombosis Res. (1979), Vol.16, pp. 245-254). For example, four tissue human serine proteases can beused in the primary bioassay, free factor Xa, prothrombinase, thrombin(IIa) and tissue plasminogen activator (tPA). The assay for tPA has beensuccessfully used before to demonstrate undesired side effects in theinhibition of the fibrinolytic process (see, e.g., J. Med. Chem. (1993),Vol. 36, pp. 314-319).

Another bioassay useful in demonstrating the utility of the compounds ofthe invention in inhibiting factor Xa demonstrates the potency of thecompounds against free factor Xa in citrated plasma. For example, theanticoagulant efficacy of the compounds of the invention will be testedusing either the prothrombin time (PT), or activated partialthromboplastin time (aPTT) while selectivity of the compounds is checkedwith the thrombin clotting time (TCT) assay. Correlation of the K_(i) inthe primary enzyme assay with the K_(i) for free factor Xa in citratedplasma will screen against compounds which interact with or areinactivated by other plasma components. Correlation of the K_(i) withthe extension of the PT is a necessary in vitro demonstration thatpotency in the free factor Xa inhibition assay translates into potencyin a clinical coagulation assay. In addition, extension of the PT incitrated plasma can be used to measure duration of action in subsequentpharmacodynamic studies.

For further information on assays to demonstrate the activity of thecompounds of the invention, see R. Lottenberg et al., Methods inEnzymology (1981), Vol. 80, pp. 341-361, and H. Ohno et al., ThrombosisResearch (1980), Vol. 19, pp. 579-588.

C. General Administration

Administration of the compounds of the invention, or theirpharmaceutically acceptable salts, in pure form or in an appropriatepharmaceutical composition, can be carried out via any of the acceptedmodes of administration or agents for serving similar utilities. Thus,administration can be, for example, orally, nasally, parenterally,topically, transdermally, or rectally, in the form of solid, semi-solid,lyophilized powder, or liquid dosage forms, such as for example,tablets, suppositories, pills, soft elastic and hard gelatin capsules,powders, solutions, suspensions, or aerosols, or the like, preferably inunit dosage forms suitable for simple administration of precise dosages.The compositions will include a conventional pharmaceutical carrier orexcipient and a compound of the invention as the/an active agent, and,in addition, may include other medicinal agents, pharmaceutical agents,carriers, adjuvants, etc.

Generally, depending on the intended mode of administration, thepharmaceutically acceptable compositions will contain about 1% to about99% by weight of a compound(s) of the invention, or a pharmaceuticallyacceptable salt thereof, and 99% to 1% by weight of a suitablepharmaceutical excipient. Preferably, the composition will be about 5%to 75% by weight of a compound(s) of the invention, or apharmaceutically acceptable salt thereof, with the rest being suitablepharmaceutical excipients.

The preferred route of administration is oral, using a convenient dailydosage regimen which can be adjusted according to the degree of severityof the disease-state to be treated. For such oral administration, apharmaceutically acceptable composition containing a compound(s) of theinvention, or a pharmaceutically acceptable salt thereof, is formed bythe incorporation of any of the normally employed excipients, such as,for example, pharmaceutical grades of mannitol, lactose, starch,pregelatinized starch, magnesium stearate, sodium saccharine, talcum,cellulose ether derivatives, glucose, gelatin, sucrose, citrate, propylgallate, and the like. Such compositions take the form of solutions,suspensions, tablets, pills, capsules, powders, sustained releaseformulations and the like.

Preferably such compositions will take the form of capsule, caplet ortablet and therefore will also contain a diluent such as lactose,sucrose, dicalcium phosphate, and the like; a disintegrant such ascroscarmellose sodium or derivatives thereof; a lubricant such asmagnesium stearate and the like; and a binder such as a starch, gumacacia, polyvinylpyrrolidone, gelatin, cellulose ether derivatives, andthe like.

The compounds of the invention, or their pharmaceutically acceptablesalts, may also be formulated into a suppository using, for example,about 0.5% to about 50% active ingredient disposed in a carrier thatslowly dissolves within the body, e.g., polyoxyethylene glycols andpolyethylene glycols (PEG), e.g., PEG 1000 (96%) and PEG 4000 (4%).

Liquid pharmaceutically administrable compositions can, for example, beprepared by dissolving, dispersing, etc., a compound(s) of the invention(about 0.5% to about 20%), or a pharmaceutically acceptable saltthereof, and optional pharmaceutical adjuvants in a carrier, such as,for example, water, saline, aqueous dextrose, glycerol, ethanol and thelike, to thereby form a solution or suspension.

If desired, a pharmaceutical composition of the invention may alsocontain minor amounts of auxiliary substances such as wetting oremulsifying agents, pH buffering agents, antioxidants, and the like,such as, for example, citric acid, sorbitan monolaurate, triethanolamineoleate, butylated hydroxytoluene, etc.

Actual methods of preparing such dosage forms are known, or will beapparent, to those skilled in this art; for example, see Remington'sPharmaceutical Sciences, 18th Ed., (Mack Publishing Company, Easton,Pa., 1990). The composition to be administered will, in any event,contain a therapeutically effective amount of a compound of theinvention, or a pharmaceutically acceptable salt thereof, for treatmentof a disease-state alleviated by the inhibition of factor Xa inaccordance with the teachings of this invention.

The compounds of the invention, or their pharmaceutically acceptablesalts, are administered in a therapeutically effective amount which willvary depending upon a variety of factors including the activity of thespecific compound employed; the metabolic stability and length of actionof the compound; the age, body weight, general health, sex, and diet ofthe patient; the mode and time of administration; the rate of excretion;the drug combination; the severity of the particular disease-states; andthe host undergoing therapy. Generally, a therapeutically effectivedaily dose is from about 0.14 mg to about 14.3 mg/kg of body weight perday of a compound of the invention, or a pharmaceutically acceptablesalt thereof; preferably, from about 0.7 mg to about 10 mg/kg of bodyweight per day; and most preferably, from about 1.4 mg to about 7.2mg/kg of body weight per day. For example, for administration to a 70 kgperson, the dosage range would be from about 10 mg to about 1.0 gram perday of a compound of the invention, or a pharmaceutically acceptablesalt thereof, preferably from about 50 mg to about 700 mg per day, andmost preferably from about 100 mg to about 500 mg per day.

Preferred Embodiments

Of the compounds of the invention as set forth above in the Summary ofthe Invention, a preferred group of compounds are those compoundswherein Z¹ is --O--, --CH₂ O-- or --S(O)_(n) -- (where n is 0 to 2); Z²is --O--, --OCH₂ -- or --S(O)_(n) -- (where n is 0 to 2); R¹ and R⁴ areeach independently hydrogen, halo, alkyl or --OR⁷ ; R² is --C(NH)NH₂,--C(NH)N(H)S(O)₂ R⁹, or --C(NH)N(H)C(O)N(H)R⁷ ; R³ is ureido, guanidino,--N(R⁷)R⁸, --N(R⁷)C(O)R⁷, (1,2)-tetrahydropyrimidinyl (optionallysubstituted by alkyl), (1,2)-imidazolyl (optionally substituted byalkyl), or (1,2)-imidazolinyl (optionally substituted by alkyl); each R⁵are independently hydrogen, halo, alkyl or haloalkyl; each R⁶ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), aralkyl (wherein the aryl is optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), heterocyclyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), orheterocyclylalkyl (wherein the heterocyclyl is optionally substituted byhalo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl); each R⁷and R⁸ is independently hydrogen, alkyl, aryl (optionally substituted byhalo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), oraralkyl (wherein the aryl is optionally substituted by halo, haloalkyl,alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino, dialkylamino,monoalkylamino, nitro, carboxy, alkoxycarbonyl, aminocarbonyl,monoalkylaminocarbonyl, or dialkylaminocarbonyl); and each R⁹ is alkyl,aryl (optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), or aralkyl (wherein the aryl is optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).

Of this group of compounds, a preferred subgroup of compounds are thosecompounds wherein Z¹ is --O--; Z² is --O--; R¹ is hydrogen or --OR⁷ ; R²is --C(NH)NH₂ ; R³ is (1,2)-tetrahydropyrimidinyl (optionallysubstituted by alkyl), (1,2)-imidazolyl (optionally substituted byalkyl), or (1,2)-imidazolinyl (optionally substituted by alkyl); R⁴ ishydrogen; each R⁵ is hydrogen or halo; and each R⁶ is aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), aralkyl (wherein the aryl is optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), heterocyclyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), orheterocyclylalkyl (wherein the heterocyclyl is optionally substituted byhalo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl); andeach R⁷ is independently hydrogen or alkyl.

Of this subgroup of compounds, a preferred class of compounds are thosecompounds wherein Z¹ is --O--; Z² is --O--; R¹ is --OR⁷ ; R² is--C(NH)NH₂ ; R³ is (1,2)-imidazolyl (optionally substituted by methyl)or (1,2)-imidazolinyl (optionally substituted by methyl); R⁴ ishydrogen; R⁵ is hydrogen; and R⁶ is aryl (optionally substituted byhalo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl); and R⁷is hydrogen or alkyl.

Of this class of compounds, a preferred subclass of compounds are thosecompounds wherein the compound is selected from the formula (I):##STR5##

Of this subclass of compounds, preferred compounds are those compoundswherein R¹ is hydroxy; R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).

Of these preferred compounds, a more preferred compound is4-hydroxy-3-[[6-(3-(1-methylimidazolin-2-yl)phenoxy)-2-(phenyl)pyrimidin-4-yl]oxy]benzamidine.

Of the class of compounds described above, another preferred subclass ofcompounds are those compounds selected from formula (II): ##STR6##

Of this subclass of compounds, preferred compounds are those compoundswherein R¹ is hydroxy; R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).

Of these preferred compounds, a more preferred compound is4-hydroxy-3-[[4-(3-(1-methylimidazolin-2-yl)phenoxy)-6-(phenyl)pyrimidin-2-yl]oxy]benzamidine.

Of the class of compounds described above, another preferred subclass ofcompounds are those compounds selected from formula (III): ##STR7##

Of this subclass of compounds, preferred compounds are those compoundswherein R¹ is hydroxy; R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).

Of these preferred compounds, a more preferred compound is4-hydroxy-3-[[2-(3-(1-methylimidazolin-2-yl)phenoxy)-6-(phenyl)pyrimidin-4-yl]oxy]benzamidine.

Preparation of Compounds of The Invention

As a matter of convenience, the following description of the preparationof the compounds of the invention is directed to the preparation ofcompounds of the invention where Z¹ and Z² are both --O--, and R² is--C(NH)NH₂. It is understood, however, that similar synthetic processesmay be used to prepare other compounds of formula (I), (II), and (III).It is also understood that in the following description, combinations ofsubstituents and/or variables (e.g., R⁷ and R⁸) on the depicted formulaeare permissible only if such combinations result in stable compounds.

Preparation of Compounds of Formula (C)

Compounds of formula (C) are intermediates in the preparation ofcompounds of the invention and are prepared as described below inReaction Scheme 1 where R⁵ and R⁶ are as described above in the Summaryof the Invention and R¹¹ is alkyl or aralkyl: ##STR8##

Compounds of formula (A) and formula (B) are commercially available, ormay be prepared according to methods known to those skilled in the art.

In general, compounds of formula (C) are prepared by first treating analkanol, preferably ethanol, to a molar excess amount of an alkalinemetal, preferably sodium. After the metal is completely dissolved, acompound of formula (A) and a compound of formula (B) are added inequimolar amounts to the solution at ambient temperature. The resultingreaction mixture is heated to reflux for about 4 to about 6 hours,preferably for about 4 hours. The solvent is removed and a strong acid,preferably hydrochloric acid, is added to the reaction mixture to afforda precipitate, which is collected and dried to afford the compound offormula (C).

Preparation of Compounds of Formula (G)

Compounds of formula (G) are intermediates in the preparation ofcompounds of the invention and are prepared as illustrated below inReaction Scheme 2 wherein R⁵ and R⁶ are as described above in theSummary of the Invention and R¹¹ is alkyl or aralkyl: ##STR9##

Compounds of formula (D) and formula (E) are commercially available, ormay be prepared according to methods known to those of ordinary skill inthe art.

In general, compounds of formula (G) are prepared by first treating analkanol, preferably ethanol, to a molar excess amount of an alkalinemetal, preferably sodium. After the metal is completely dissolved, acompound of formula (D) and a compound of formula (E) in equimolaramounts are added to the solution. The reaction mixture is heated forabout 8 to about 16 hours, preferably for about 16 hours. The solvent isremoved and the residue dissolved in water. A strong acid, preferablyhydrochloric acid, is added to the solution and the resultingprecipitate is collected and dried to form a compound of formula (G).

Preparation of Compounds of Formula (H)

Compounds of formula (H) are intermediates in the preparation of thecompounds of the invention and are prepared as illustrated below inReaction Scheme 3 wherein R⁵ and R⁶ are as described above in theSummary of the Invention: ##STR10##

Compounds of formula (C) are prepared by methods described herein.

In general, compounds of formula (H) are prepared by treating a compoundof formula (C) with a slight molar excess of a chlorinating agent,preferably phosphorus oxychloride, in the presence of a base, such asN,N-diethylaniline. The resulting reaction mixture is refluxed for about2 to about 4 hours, preferably for about 3 hours, and then ice is addedto the solution to afford a precipitate, which is collected and dried toafford a compound of formula (H).

In a similar manner, compounds of formula (G) may be chlorinated to formthe dichloro-substituted compounds.

Preparation of Compounds of Formula (Ia)

Compounds of formula (Ia) are compounds of the invention selected fromformula (I), and are prepared as illustrated below in Reaction Scheme 4where R¹, R³, R⁴, R⁵ and R⁶ are as described above in the Summary of theInvention. Prior to Step 1 compounds of formula (H), formula (J) andformula (L) wherein the R¹, R³, R⁴, R⁵ and R⁶ substituents containadditional reactive hydroxy or amino groups may be treated with theappropriate oxygen-or nitrogen-protecting group according to methodsknown to those skilled in the art, such as those described in Greene, T.W., Protective Groups in Organic Synthesis (1981), John Wiley & Sons,New York, New York. Such protected substituents will be deprotectedduring the reaction conditions of Step 3 to form the desired R¹, R³, R⁴,R⁵ and R⁶ substituents. Alternatively, compounds of formula (H), formula(J) and formula (L) may already be present in an oxygen- ornitrogen-protected form and may be deprotected by methods known to thoseskilled in the art or, as a result of the reaction conditions of Step 3,may become deprotected to form the desired substituent. ##STR11##

Compounds of formula (H) are prepared by methods disclosed herein.Compounds of formula (J) and formula (L) are commercially available ormay be prepared according to methods known to those skilled in the art.

In general, the compounds of formula (Ia) are prepared by first treatinga compound of formula (H) in an aprotic solvent, for example,acetonitrile, with an equimolar amount of a compound of formula (J) inthe presence of a base, for example, cesium carbonate, at temperaturesbetween about 20° C. and 120° C., preferably at ambient temperature, fora period of time sufficient to complete the desired reaction asmonitored by thin layer chromatography (TLC). The compound of formula(K) is then isolated from the reaction mixture by standard isolationtechniques, such as extraction, in vacuo removal of solvent, and flashchromatography.

The compound of formula (K) in an aprotic solvent, for example, DMSO, isthen treated with an equimolar amount of a compound of formula (L) inthe presence of a base, for example, cesium carbonate, at temperaturesbetween about 20° C. and 120° C., preferably at about 50° C., for aperiod of time sufficient to complete the desired reaction, for example,for about 24 hours. The reaction mixture is cooled to ambienttemperature and the compound of formula (M) is then isolated from thereaction mixture through standard isolation techniques, such asextraction, in vacuo removal of solvent, and flash chromatography.

The compound of formula (M) is dissolved in an anhydrous alkanol,preferably ethanol, and then anhydrous mineral acid, preferably HCl, isadded to the solution over a period of time sufficient to saturate theacid into the solution while maintaining the reaction temperatures atabout -78° C. After saturation is complete, the reaction vessel issealed and the reaction mixture is allowed to warm to ambienttemperature and stirred between 12 and 24 hours, preferably for about 18hours. The solvent is removed in vacuo and the resulting residue isdissolved in fresh anhydrous alkanol, preferably ethanol, and thentreated with anhydrous ammonia (gas) at temperatures from betweenambient temperature and about 100° C. from about 1 to about 48 hours,preferably at about 60° C. and for about 2 hours. The compound offormula (Ia) is then isolated from the reaction mixture by standardisolation techniques, for example, in vacuo removal of solvent andpurification by high performance liquid chromatography (HPLC). Duringthis last step, compounds of formula (G) where any R¹, R³, R⁴, R⁵, or R⁶substituent is in an oxygen- or nitrogen-protected form are deprotectedto form compounds of formula (Ia) where R¹, R³, R⁴, R⁵ or R⁶ is asdefined above in the Summary of the Invention.

Alternatively, instead of treating the resulting residue above withanhydrous ammonia (gas), the resulting residue may be treated with acompound of the formula NH₂ OR⁷ to afford the corresponding compound offormula (Ia) wherein R² is --C(NH)N(H)OR⁷.

Compounds of formula (Ia) wherein R³ is --C(NH)NH₂ or --C(NH)N(H)OR⁷ areproduced from the corresponding cyano compounds in a similar manner asthat described above for compound of formula (M).

Compounds of formula (Ia) wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹contains an alkoxycarbonyl group or a --C(O)OR⁷ group where R⁷ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl) may be prepared from the corresponding activatedacid, such as an acid halide by techniques known to those of ordinaryskill in the art.

Compounds of formula (Ia) wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹contains an aminocarbonyl group, a monoalkylaminocarbonyl group, adialkylaminocarbonyl group, a --C(O)N(R⁷)R⁸ group or a --C(O)OR⁷ group(where each R⁷ or R⁸ is independently alkyl, aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl) or aralkyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkyl, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl) may alsobe hydrolyzed under acidic conditions to prepare corresponding compoundsof the invention where R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹ contains acarboxy or a --C(O)OH group.

Compounds of formula (Ia) wherein R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹contains a carboxy group, an alkoxycarbonyl group, or a --C(O)OR⁷ groupwhere R⁷ is hydrogen, alkyl, aryl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl), oraralkyl (optionally substituted by halo, haloalkyl, alkyl, aryl,aralkyl, hydroxy, alkoxy, aralkyl, amino, dialkylamino, monoalkylamino,nitro, carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl,or dialkylaminocarbonyl) may also be amidated under standard amidationconditions to form the corresponding compounds of formula (Ia) where R¹,R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹ contains an aminocarbonyl group, amonoalkylaminocarbonyl group, a dialkylaminocarbonyl group or a--C(O)N(R⁷)R⁸ group where R⁷ and R⁸ are independently hydrogen, alkyl,aryl (optionally substituted by halo, haloakyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), or aralkyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkyl, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl).

Compounds of formula (Ia) where R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸ or R⁹contains a nitro group may also be reduced under standard conditions toproduce the corresponding compounds of formula (Ia) where R¹, R³, R⁴,R⁵, R⁶, R⁷, R⁸ or R⁹ contains an amino group, which may also be treatedwith the appropriate alkylating agents or acylating agents to afford thecorresponding compounds of formula (Ia) where R¹, R³, R⁴, R⁵, R⁶, R⁷, R⁸or R⁹ contains a monoalkylamino group, a dialkylamino group, a --N(R⁷)R⁸group or a --N(R⁷)C(O)R⁷ where each R⁷ and R⁸ is independently hydrogen,alkyl, or aralkyl (optionally substituted by halo, haloalkyl, alkyl,aryl, aralkyl, hydroxy, alkoxy, aralkyl, amino, dialkylamino,monoalkylamino, nitro, carboxy, alkoxycarbonyl, aminocarbonyl,monoalkylaminocarbonyl, or dialkylaminocarbonyl).

Compounds of formula (Ia) may also be further treated with theappropriate acid halide, preferably acid chloride, or with theappropriate acid anhydride or an equivalent, to yield compounds of theinvention wherein R² is --C(NH)N(H)C(O)R⁷ where R⁷ is hydrogen, alkyl,aryl (optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl), or aralkyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkyl, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl).

Alternatively, compounds of formula (Ia) may further be treated with acompound of formula Cl--C(O)--OR⁹ or its functional equivalent to yieldcompounds of the invention where R² is --C(NH)N(H)C(O)OR⁹ where R⁹ isalkyl, aryl (optionally substituted by halo, haloalkyl, alkyl, aryl,aralkyl, hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino,nitro, carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl,or dialkylaminocarbonyl), or aralkyl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkyl, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl).

Alternatively, compounds of formula (Ia) may also be further treatedwith compounds of the formula R⁹ --S(O)₂ -imidazole (where R⁹ is definedabove in the Summary of the Invention) in a polar solvent, such asmethylene chloride, at ambient temperature to afford compounds of theinvention where R² is --C(NH)N(H)S(O)₂ R⁹ where R⁹ is defined above inthe Summary of the Invention.

Alternatively, compounds of formula (Ia) may be further treated with anappropriately N--R⁷ -substituted phenylcarbamate in a polar solvent,preferably methylene chloride, at ambient temperature, for about 6 to 24hours, preferably for about 12 hours, to afford compounds of theinvention where R² is --C(NH)N(H)C(O)N(H)R⁷ where R⁷ is defined above inthe Summary of the Invention.

Compounds of formula (Ia) which contain an unoxidized sulfur atom may beoxidized with the appropriate oxidizing agent to produce compoundscontaining oxidized sulfur (i.e., --S(O)_(n) -- where n is 1 or 2).

In a similar manner to the reactions described above, compounds offormula (II) and (III) may be prepared.

All compounds of the invention as prepared above which exist in freebase or acid form may be converted to their pharmaceutically acceptablesalts by treatment with the appropriate inorganic or organic base oracid. Salts of the compounds prepared above may be converted to theirfree base or acid form by standard techniques.

The following specific preparations and examples are provided as a guideto assist in the practice of the invention, and are not intended as alimitation on the scope of the invention.

PREPARATION 1 Compounds of Formula (C)

A. To ethanol (200 mL) was added sodium (7.9 g, 34 mmol). Afterdissolution, benzamidine hydrochloride (20.0 g, 0.128 mmol) and diethylmalonate (20 mL, 132 mmol) were added. The reaction was heated for 4hours and the solvent was removed in vacuo. Hydrochloric acid was addedand a precipitate formed. The solid was collected and dried in a vacuumoven to give 10.8 g (45%) of 2-phenyl-4,6-dihydroxypyrimidine.

B. In a similar manner, the following compounds of formula (C) are made:

2-(4-methylphenyl)-4,6-dihydroxypyrimidine;

2-(3-chlorophenyl)-4,6-dihydroxypyrimidine;

2-(4-phenylphenyl)-4,6-dihydroxypyrimidine;

2-(4-(benzyloxy)phenyl)-4,6-dihydroxypyrimidine;

2-(4-methoxyphenyl)-4,6-dihydroxypyrimidine;

2-(4-(benzyloxycarbonylamino)phenyl)-4,6-dihydroxypyrimidine;

2-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-4,6-dihydroxypyrimidine;

2-(4-(dimethylamino)phenyl)-4,6-dihydroxypyrimidine;

2-(4-(aminocarbonyl)phenyl)-4,6-dihydroxypyrimidine;

2-(1-(benzyloxycarbonyl)piperidin-4-yl)-4,6-dihydroxypyrimidine;

2-(1-ethylpiperidin-4-yl)-4,6-dihydroxypyrimidine;

2-(pyridin-2-yl)-4,6-dihydroxypyrimidine;

2-(imidazolin-2-yl)-4,6-dihydroxypyrimidine; and

2-(piperidin-1-yl)methyl-4,6-dihydroxypyrimidine.

C. In a manner similar to that described above, other compounds offormula (C) may be prepared.

PREPARATION 2 Compounds of Formula (G)

A. To ethanol (100 mL) was added sodium (3.3 g, 14 mmol). Afterdissolution, thiourea (10.7 g, 14 mmol) and ethyl benzoylacetate (24 mL,14 mmol) were added. The reaction was heated for 16 hours and thesolvent was removed in vacuo. Water was added to dissolve the residueand hydrochloric acid was added and a precipitate formed. The solid wascollected and dried in a vacuum oven to give 16 g (56%) of2-mercapto-4-hydroxy-6-phenylpyrimidine.

B. 2-Mercapto-4-hydroxy-6-phenylpyrimidine (8.6 g, 42 mmol) andchloroacetic acid (8.5 g, 90 mmol) were slurried in water (100 mL).After refluxing the mixture for 24 hours, hydrochloric acid was addedand the precipitate was isolated by filtration. The solid was dried in avacuum oven to give 6.6 g (83%) of 2,4-dihydroxy-6-phenylpyrimidine.

C. In a similar manner, the following compounds of formula (G) are made:

6-(4-methylphenyl)-2,4-dihydroxypyrimidine;

6-(3-chlorophenyl)-2,4-dihydroxypyrimidine;

6-(4-phenylphenyl)-2,4-dihydroxypyrimidine;

6-(4-(benzyloxy)phenyl)-2,4-dihydroxypyrimidine;

6-(4-methoxyphenyl)-2,4-dihydroxypyrimidine;

6-(4-(benzyloxycarbonylamino)phenyl)-2,4-dihydroxypyrimidine;

6-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-2,4-dihydroxypyrimidine;

6-(4-(dimethylamino)phenyl)-2,4-dihydroxypyrimidine;

6-(4-(aminocarbonyl)phenyl)-2,4-dihydroxypyrimidine;

6-(1-(benzyloxycarbonyl)piperidin-4-yl)-2,4-dihydroxypyrimidine;

6-(1-ethylpiperidin-4-yl)-2,4-dihydroxypyrimidine;

6-(pyridin-2-yl)-2,4-dihydroxypyrimidine;

6-(imidazolin-2-yl)-2,4-dihydroxypyrimidine; and

6-(piperidin-1-yl)methyl-2,4-dihydroxypyrimidine.

D. In a manner similar to that described above in Paragraphs A and B,other compounds of formula (G) may be prepared.

PREPARATION 3 Compounds of Formula (H)

A. To 4,6-dihydroxy-2-phenylpyrimidine (10.8 g, 57 mmol) was addedphosphorous oxychloride (58 mL, 62 mmol) and N,N-diethylaniline (16 mL,10 mmol). After refluxing for 3 hours, ice was added to the solution anda precipitate formed. The solid was collected and dried in a vacuum ovento give 11.4 g (88%) of 4,6-dichloro-2-phenylpyrimidine; m.p. 94-95° C.

B. In a similar manner, the following compound was made:

2,4-dichloro-6-phenylpyrimidine; m.p. 85-87° C.

C. In a similar manner, the following compounds of formula (H) are made:

2-(4-methylphenyl)-4,6-dichloropyrimidine;

2-(3-chlorophenyl)-4,6-dichloropyrimidine;

2-(4-phenylphenyl)-4,6-dichloropyrimidine;

2-(4-(benzyloxy)phenyl)-4,6-dichloropyrimidine;

2-(4-methoxyphenyl)-4,6-dichloropyrimidine;

2-(4-(benzyloxycarbonylamino)phenyl)-4,6-dichloropyrimidine;

2-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-4,6-dichloropyrimidine;

2-(4-(dimethylamino)phenyl)-4,6-dichloropyrimidine;

2-(4-(aminocarbonyl)phenyl)-4,6-dichloropyrimidine;

2-(1-(benzyloxycarbonyl)piperidin-4-yl)-4,6-dichloropyrimidine;

2-(1-ethylpiperidin-4-yl)-4,6-dichloropyrimidine;

2-(pyridin-2-yl)-4,6-dichloropyrimidine;

2-(imidazolin-2-yl)-4,6-dichloropyrimidine; and

2-(piperidin-1-yl)methyl-4,6-dichloropyrimidine.

D. In a manner similar to that described above in Paragraphs A, thefollowing intermediates of the compounds of the invention are prepared:

6-(4-methylphenyl)-2,4-dichloropyrimidine;

6-(3-chlorophenyl)-2,4-dichloropyrimidine;

6-(4-phenylphenyl)-2,4-dichloropyrimidine;

6-(4-(benzyloxy)phenyl)-2,4-dichloropyrimidine;

6-(4-methoxyphenyl)-2,4-dichloropyrimidine;

6-(4-(benzyloxycarbonylamino)phenyl)-2,4-dichloropyrimidine;

6-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-2,4-dichloropyrimidine;

6-(4-(dimethylamino)phenyl)-2,4-dichloropyrimidine;

6-(4-(aminocarbonyl)phenyl)-2,4-dichloropyrimidine;

6-(1-(benzyloxycarbonyl)piperidin-4-yl)-2,4-dichloropyrimidine;

6-(1-ethylpiperidin-4-yl)-2,4-dichloropyrimidine;

6-(pyridin-2-yl)-2,4-dichloropyrimidine;

6-(imidazolin-2-yl)-2,4-dichloropyrimidine; and

6-(piperidin-1-yl)methyl-2,4-dichloropyrimidine.

E. In a similar manner, other intermediates of the compounds of theinvention may be prepared.

PREPARATION 4 Compounds of Formula (K)

A. To 4,6-dichloro-2-phenylpyrimidine (1.5 g, 6.7 mmol) in acetonitrile(20 mL) was added Cs₂ CO₃ (2.4 g, 7.4 mmol) and3-hydroxy4-(benzyloxy)benzonitrile (1.5 g, 6.5 mmol). After stirring for24 hours, water was added to the reaction mixture and the resultingsolid was collected by suction filtration to give 2.6 g of3-[(6-chloro-2-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

B. In a similar manner, the following intermediates of compounds of theinvention were made:

3-[(6-chloro-4-phenylpyrimidin-2-yl)oxy]-4-(benzyloxy)benzonitrile; and

3-[(2-chloro-6-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

C. In a similar manner, the following compounds of formula (K) are made:

3-[(2-(4-methylphenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(3-chlorophenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-phenylphenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(benzyloxy)phenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-methoxyphenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(benzyloxycarbonylamino)phenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(dimethylamino)phenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(aminocarbonyl)phenyl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(1-(benzyloxycarbonyl)piperidin-4-yl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(1-ethylpiperidin-4-yl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(pyridin-2-yl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(imidazolin-2-yl)-6-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;and

3-[(2-(piperidin-1-yl)methyl-6-chloropyrimidinyl)oxy]-4-(benzyloxy)benzonitrile.

D. In a similar manner to that described above, the followingintermediates of the compounds of the invention are made:

3-[(6-(4-methylphenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(3-chlorophenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-phenylphenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(benzyloxy)phenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-methoxyphenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(benzyloxycarbonylamino)phenyl)-2-chloropyrimidinyl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(dimethylamino)phenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(aminocarbonyl)phenyl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(1-(benzyloxycarbonyl)piperidin-4-yl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(1-ethylpiperidin-4-yl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(pyridin-2-yl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(imidazolin-2-yl)-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;and

3-[(6-(piperidin-I-yl)methyl-2-chloropyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

E. In a similar manner to that described above, other intermediates ofthe compounds of the invention may be prepared.

PREPARATION 5 Compounds of Formula (M)

A. To 3-[(6-chloro-2-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile(1.0 g, 2.4 mmol) in DMSO (12 mL) was added Cs₂ CO₃ (0.8 g, 2.5 mmol)and 2-(3-hydroxyphenyl)-1-methylimidazoline (0.44 g, 2.5 mmol). Afterstirring in an oil bath at 50° C. for 24 hours, the reaction waspartitioned with water and ethyl acetate. The layers were separated,washed with water and brine, dried (Na₂ SO₄), and the solvent wasremoved in vacuo. The residue was chromatographed on silica gel with CH₂CL₂ /MeOH/NH₄ OH (120/5/1) to give 0.5 g of3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

B. In a similar manner, the following compounds of formula (M) weremade:

3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-4-phenylpyrimidin-2-yl)oxy]-4-(benzyloxy)benzonitrile;and

3-[(2-[3-(1-methylimidazolin-2-yl)phenoxy]-6-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

C. In a similar manner, the following compounds of formula (M) are made:

3-[(2-(4-methylphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(3-chlorophenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-phenylphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(benzyloxy)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-methoxyphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(benzyloxycarbonylamino)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(dimethylamino)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(4-(aminocarbonyl)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(1-(benzyloxycarbonyl)piperidin-4-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(1-ethylpiperidin-4-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(pyridin-2-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(2-(imidazolin-2-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;and

3-[(2-(piperidin-1-yl)methyl-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

D. In a manner similar to that described above, the followingintermediates of the compounds of the invention are prepared:

3-[(6-(4-methylphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(3-chlorophenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-phenylphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(benzyloxy)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-methoxyphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(benzyloxycarbonylamino)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-(benzyloxy)benzonitrile;

3-[(6-(3-((methyl)(benzyloxycarbonyl)amino)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(dimethylamino)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(4-(aminocarbonyl)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(1-(benzyloxycarbonyl)piperidin-4-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(1-ethylpiperidin-4-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(pyridin-2-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;

3-[(6-(imidazolin-2-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile;and

3-[(6-(piperidin-1-yl)methyl-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile.

E. In a similar manner to that described above, other intermediates ofthe compounds of the invention may be prepared.

EXAMPLE 1 Compounds of Formula (I), (II), and (III)

A. To3-[(6-[3-(I-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]-4-(benzyloxy)benzonitrile(0.4 g, 0.7 mmol) dissolved in ethanol (15 mL) and cooled in a dryice/isopropanol bath was bubbled HCl (g). After the solution wassaturated the reaction flask was sealed and allowed to warm to ambienttemperature and stirred for 18 hours. The solvent was removed in vacuoand the residue was triturated with ether. The ether was decanted offand the residue was dissolved in ethanol (6 mL). The solution was cooledin a dry ice/isopropanol bath and ammonia (g) was bubbled into thesolution. The reaction flask was sealed and heated in an oil bath at 60°C. for 2 hours. The solvent was removed in vacuo and the residue waspurified by HPLC on a C18 Dynamax column with a 10-40% acetonitrile inwater gradient with 0.1% trifluoroacetic acid to give4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine,trifluoroacetic acid salt, as a pure trifluoroacetic acid salt; NMR(DMSO-d₆) 11.2 (s, 1), 10.4 (s, 1), 9.1 (s, 2), 8.9 (s, 2), 7.8 (m, 7),7.4 (m, 4), 7.2 (d, 1), 6.7 (s, 1), 4.1 (m, 2), 4.0 (m, 2), 3.1 (s, 3)ppm.

B. In a similar manner, the following compounds of the invention weremade:

4-hydroxy-3-[(4-[3-(1-methylimidazolin-2-yl)phenoxy]-6-phenylpyrimidin-2-yl)oxy]benzamidineand4-hydroxy-3-[(2-[3-(1-methylimidazolin-2-yl)phenoxy]-6-phenylpyrimidin-4-yl)oxy]benzamidine,trifluoroacetic acid salt, NMR (DMSO-d₆) 10.4 (s, 1), 9.1 (m, 2), 8.8(m, 2), 8.0 (d, 2), 7.4-7.8 (m, 11), 7.1 (m, 1), 4.0 (m, 4), 3.1 (s, 3)ppm.

C. In a similar manner, the following compounds of the invention aremade:

3-[(2-(4-methylphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(3-chlorophenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-phenylphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-hydroxyphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-methoxyphenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-aminophenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(3-(methylamino)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-(dimethylamino)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(4-(aminocarbonyl)phenyl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(piperidin-4-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(1-ethylpiperidin-4-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(pyridin-2-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(imidazolin-2-yl)-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(2-(piperidin-1-yl)methyl-6-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-methylphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(3-chlorophenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-phenylphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-hydroxyphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-methoxyphenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-aminophenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(3-(methylamino)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-(dimethylamino)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(4-(aminocarbonyl)phenyl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(piperidin-4-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(1-ethylpiperidin-4-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(pyridin-2-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;

3-[(6-(imidazolin-2-yl)-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine;and

3-[(6-(piperidin-1-yl)methyl-2-[3-(1-methylimidazolin-2-yl)phenoxy]pyrimidin-4-yl)oxy]-4-hydroxybenzamidine.

D. In a manner similar to that described above in Paragraph A, othercompounds of the invention may be prepared.

EXAMPLE 2

This example illustrates the preparation of representativepharmaceutical compositions for oral administration containing acompound of the invention, or a pharmaceutically acceptable saltthereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        A.        Ingredients     % wt./wt.                                           ______________________________________                                                Compound of the invention                                                                   20.0%                                                           Lactose       79.5%                                                           Magnesium stearate                                                                           0.5%                                                   ______________________________________                                    

The above ingredients are mixed and dispensed into hard-shell gelatincapsules containing 100 mg each, one capsule would approximate a totaldaily dosage.

    ______________________________________                                        B.        Ingredients     % wt./wt.                                           ______________________________________                                                Compound of the invention                                                                   20.0%                                                           Magnesium stearate                                                                           0.9%                                                           Starch         8.6%                                                           Lactose       69.6%                                                           PVP (polyvinylpyrrolidine)                                                                   0.9%                                                   ______________________________________                                        ______________________________________                                    

The above ingredients with the exception of the magnesium stearate arecombined and granulated using water as a granulating liquid. Theformulation is then dried, mixed with the magnesium stearate and formedinto tablets with an appropriate tableting machine.

    ______________________________________                                        C.        Ingredients                                                         ______________________________________                                                Compound of the invention                                                                   0.1     g                                                       Propylene glycol                                                                            20.0    g                                                       Polyethylene glycol 400                                                                     20.0    g                                                       Polysorbate 80                                                                              1.0     g                                                       Water         q.s. 100                                                                              mL                                              ______________________________________                                    

The compound of the invention is dissolved in propylene glycol,polyethylene glycol 400 and polysorbate 80. A sufficient quantity ofwater is then added with stirring to provide 100 mL of the solutionwhich is filtered and bottled.

    ______________________________________                                        D.        Ingredients     % wt./wt.                                           ______________________________________                                                Compound of the invention                                                                   20.0%                                                           Peanut Oil    78.0%                                                           Span 60        2.0%                                                   ______________________________________                                    

The above ingredients are melted, mixed and filled into soft elasticcapsules.

    ______________________________________                                        E.      Ingredients        % wt./wt.                                          ______________________________________                                        Compound of the invention                                                                            1.0%                                                   Methyl or carboxymethyl cellulose                                                                    2.0%                                                   0.9% saline            q.s. 100 mL                                            ______________________________________                                    

The compound of the invention is dissolved in the cellulose/salinesolution, filtered and bottled for use.

EXAMPLE 3

This example illustrates the preparation of a representativepharmaceutical formulation for parenteral administration containing acompound of the invention, or a pharmaceutically acceptable saltthereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        Ingredients                                                                   ______________________________________                                        Compound of the invention                                                                       0.02         g                                              Propylene glycol  20.0         g                                              Polyethylene glycol 400                                                                         20.0         g                                              Polysorbate 80    1.0          g                                              0.9% Saline solution                                                                            q.s. 100     mL                                             ______________________________________                                    

The compound of the invention is dissolved in propylene glycol,polyethylene glycol 400 and polysorbate 80. A sufficient quantity of0.9% saline solution is then added with stirring to provide 100 mL ofthe I.V. solution which is filtered through a 0.2 m membrane filter andpackaged under sterile conditions.

EXAMPLE 4

This example illustrates the preparation of a representativepharmaceutical composition in suppository form containing a compound ofthe invention, or a pharmaceutically acceptable salt thereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        Ingredients        % wt./wt.                                                  ______________________________________                                        Compound of the invention                                                                         1.0%                                                      Polyethylene glycol 1000                                                                         74.5%                                                      Polyethylene glycol 4000                                                                         24.5%                                                      ______________________________________                                    

The ingredients are melted together and mixed on a steam bath, andpoured into molds containing 2.5 g total weight.

EXAMPLE 5

This example illustrates the preparation of a representativepharmaceutical formulation for insufflation containing a compound of theinvention, or a pharmaceutically acceptable salt thereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        Ingredients           % wt./wt.                                               ______________________________________                                        Micronized compound of the invention                                                                 1.0%                                                   Micronized lactose    99.0%                                                   ______________________________________                                    

The ingredients are milled, mixed, and packaged in an insufflatorequipped with a dosing pump.

EXAMPLE 6

This example illustrates the preparation of a representativepharmaceutical formulation in nebulized form containing a compound ofthe invention, or a pharmaceutically acceptable salt thereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        Ingredients        % wt./wt.                                                  ______________________________________                                        Compound of the invention                                                                         0.005%                                                    Water              89.995%                                                    Ethanol            10.000%                                                    ______________________________________                                    

The compound of the invention is dissolved in ethanol and blended withwater. The formulation is then packaged in a nebulizer equipped with adosing pump.

EXAMPLE 7

This example illustrates the preparation of a representativepharmaceutical formulation in aerosol form containing a compound of theinvention, or a pharmaceutically acceptable salt thereof, e.g.,4-hydroxy-3-[(6-[3-(1-methylimidazolin-2-yl)phenoxy]-2-phenylpyrimidin-4-yl)oxy]benzamidine:

    ______________________________________                                        Ingredients        % wt./wt.                                                  ______________________________________                                        Compound of the invention                                                                         0.10%                                                     Propellant 11/12   98.90%                                                     Oleic acid         1.00%                                                      ______________________________________                                    

The compound of the invention is dispersed in oleic acid and thepropellants. The resulting mixture is then poured into an aerosolcontainer fitted with a metering valve.

EXAMPLE 8 In Vitro Assay for Factor Xa and Thrombin

This assay demonstrates the activity of the compounds of the inventiontowards factor Xa, thrombin and tissue plasminogen activator. Theactivities were determined as an initial rate of cleavage of the peptidep-nitroanilide by the enzyme. The cleavage product, p-nitroaniline,absorbs at 405 nm with a molar extinction coefficient of 9920 M⁻¹ cm⁻¹.

Reagents and Solutions

Dimethyl sulfoxide (DMSO) (Baker analyzed grade).

Assay buffer:

50 mM Tris HCl, 150 mM NaCl, 2.5 mM CaCl₂, and

0.1% polyethylene glycol 6000, pH 7.5.

Enzymes (Enzyme Research Lab.):

1. Human factor Xa stock solution: 0.281 mg/mL in assay buffer, storedat -80° C. (working solution (2×): 106 ng/mL or 2 nM in assay buffer,prepare prior to use).

2. Human thrombin stock solution: Concentration as specified by thesupplier, stored at -80° C. (working solution (2×): 1200 ng/mL or 32 nMin assay buffer, prepare prior to use).

3. Human tissue plasminogen activator (tPA) (Two chains, Sigma orAmerican Diagnostica Inc.) stock solution: Concentration as specified bythe supplier, stored at -80° C. (working solution (2×): 1361 ng/mL or 20nM in assay buffer, prepare prior to use).

Chromogenic substrates (Pharmacia Hepar Inc.):

1. S2222 (FXa assay) stock solution: 6 mM in deionized H₂ O, store at 4°C. (working solution (4×): 656 μM in assay buffer).

2. S2302 (Thrombin assay) stock solution: 10 mM in deionized H₂ O,stored at 4° C. (working solution (4×): 1200 μM in assay buffer).

3. S2288 (tPA assay) stock solution: 10 mM in deionized H₂ O, stored at4° C. (working solution (4×): 1484 μM in assay buffer for Sigma tPA, or1120 μM for American Diagnostica tPA).

Standard inhibitor compound stock solution:

5 mM in DMSO, stored at -20° C.

Test compounds (compounds of the invention) stock solutions:

10 mM in DMSO, stored at -20° C.

Assay Procedure

Assays were performed in 96-well microtiter plates in a total volume of200 I. Assay components were in final concentration of 50 mM Tris HCl,150 mM NaCl, 2.5 mM CaCl₂, 0.1% polyethylene glycol 6000, pH 7.5, in theabsence or presence of the standard inhibitor or the test compounds andenzyme and substrate at following concentrations: (1) 1 nM factor Xa(0.1 nM or 0.2 nM factor Xa for compounds with K_(i) Xa in low picomolarrange) and 164 μM S2222; (2) 16 nM thrombin and 300 μM S2302; and (3) 10nM tPA and 371 μM or 280 μM S2288. Concentrations of the standardinhibitor compound in the assay were from 5 μM to 0.021 μM in 1 to 3dilution. Concentration of the test compounds in the assay typicallywere from 10 μM to 0.041 μM in 1 to 3 dilution. For potent testcompounds, the concentrations used in the factor Xa assay were furtherdiluted 100 fold (100 nM to 0.41 nM) or 1000 fold (10 nM to 0.041 nM).All substrate concentrations used are equal to their Km values under thepresent assay conditions. Assays were performed at ambient temperature.

The first step in the assay was the preparation of 10 mM test compoundstock solutions in DMSO (for potent test compounds, 10 mM stocksolutions were further diluted to 0.1 or 0.01 μM for the factor Xaassay), followed by the preparation of test compound working solutions(4×) by a serial dilutions of 10 mM stock solutions with Biomek 1000 in96 deep well plates as follows:

(a) Prepare a 40 μM working solution by diluting the 10 mM stock 1 to250 in assay buffer in 2 steps: 1 to 100, and 1 to 2.5.

(b) Make another five serial dilutions (1:3) of the 40 μM solution (600μL for each concentration). A total of six diluted test compoundsolutions were used in the assay.

Standard inhibitor compound (5 mM stock) or DMSO (control) went throughthe same dilution steps as those described above for test compounds.

The next step in the assay was to dispense 50 ,L of the test compoundworking solutions (4×) (from 40 μM to 0.164 μM) in duplicate tomicrotiter plates with Biomek. To this was added 100 μL of enzymeworking solution (2×) with Biomek. The resulting solutions wereincubated at ambient temperature for 10 minutes.

To the solutions was added 50 μL of substrate working solution (4×) withBiomek.

The enzyme kinetics were measured at 405 nm at 10 seconds intervals forfive minutes in a THERMOmax plate reader at ambient temperature. When alower concentration of factor Xa was needed in the factor Xa assay, theenzyme kinetics were measured for fifteen minutes (0.2 nM factor Xa) orthirty minutes (0.1 nM factor Xa) at ambient temperature.

Calculation of K_(i) of the Test Compounds

Enzyme initial rates were calculated as mOD/min based on the first twominutes readings. The IC₅₀ values were determined by fitting the data tothe log-logit equation (linear) or the Morrison equation (non-linear)with an EXCEL spread-sheet. K_(i) values were then obtained by dividingthe IC₅₀ by 2. Routinely, K_(i) (factor Xa) values less than 3 nM werecalculated from the Morrison equation.

Compounds of the invention, when tested in this assay, demonstrated theselective ability to inhibit human factor Xa and human thrombin.

EXAMPLE 9 In Vitro Assay for Human Prothrombinase

This assay demonstrates the ability of the compounds of the invention toinhibit prothrombinase. Prothrombinase (PTase) catalyzes the activationof prothrombin to yield fragment 1.2 plus thrombin with meizothrombin asthe intermediate. This assay is an end point assay. Activity of theprothrombinase is measured by activity of thrombin (one of the reactionproducts) or by the amount of thrombin formed/time based on a thrombinstandard curve (nM vs mOD/min). For determination of IC₅₀ (PTase) of thecompounds of the invention, PTase activity was expressed by thrombinactivity (mOD/min).

Materials

Enzymes:

1. Human factor Va (Haematologic Technologies Inc., Cat# HCVA-01 10)working solution: 1.0 mg/mL in 50% glycerol, 2 mM CaCl₂, stored at -20°C.

2. Human factor Xa (Enzyme Res. Lab. cat# HFXa1011) working solution:0.281 mg/mL in assay buffer (without BSA), stored at -80° C.

3. Human prothrombin (FII) (Enzyme Res. Lab., Cat# HP1002) workingsolution:

Diluted FII to 4.85 mg/mL in assay buffer (without BSA), stored at -80°C.

Phospholipid (PCPS) vesicles:

PCPS vesicles (80% PC, 20% PS) were prepared by modification of themethod reported by Barenholz et al., Biochemistry (1977), Vol. 16, pp.2806-2810.

Phosphatidyl serine (Avanti Polar Lipids, Inc., Cat#840032):

10 mg/mL in chloroform, purified from brain, stored -20° C. undernitrogen or argon.

Phosphatidyl Choline (Avanti Polar Lipids, Inc., Cat# 850457):

50 mg/ml in chloroform, synthetic 16:0-18:1 Palmitoyl-Oleoyl, stored at-20° C. under nitrogen or argon.

Spectrozyme-TH (American Diagnostica Inc., Cat# 238L, 50 μmoles, storedat ambient temperature) working solution: Dissolved 50 μmoles in 10 mLdH₂ O.

BSA (Sigma Chem Co., Cat# A-7888, FractionV, RIA grade).

Assay buffer: 50 mM Tris HCl, pH 7.5, 150 mM NaCl, 2.5 mM CaCl₂, 0.1%PEG 6000 (BDH), 0.05% BSA (Sigma, Fr.V, RIA grade).

For one plate assay, prepare the following working solutions:

1. Prothrombinase complex:

(a) 100 μM PCPS (27.5 μL of PCPS stock (4.36 mM) diluted to final 1200μL with assay buffer.

(b) 25 nM Human factor Va: 5.08 μL of Va stock(1 mg/mL) was diluted tofinal 1200 μL with assay buffer.

(c) 5 pM Human factor Xa: Dilute factor Xa stock (0.281 mg/mL)1:1,220,000 with assay buffer. Prepare at least 1200 μL.

Combine equal volumes (1100 μL) of each component in the order of PCPS,Va and Xa. Use immediately or store in ice (bring to ambient temperaturebefore use).

2. 6 μM Human prothrombin (FII): dilute 124 μL of FII stock (4.85 mg/mL)to final 1400 μL with assay buffer.

3. 20 mM EDTA/Assay buffer: 0.8 mL of 0.5M EDTA (pH 8.5) plus 19.2 mLassay buffer.

4. 0.2 mM Spectrozyme-TH/EDTA buffer: 0.44 mL of SPTH stock (5 mM) plus10.56 mL of 20 mM EDTA/assay buffer.

5. Test compounds (compounds of the invention):

Prepare a working solution (5×) from 10 mM stock (DMSO) and make aseries of 1:3 dilution. Compounds were assayed at 6 concentrations induplicate.

Assay Conditions and Procedure

Prothrombinase reaction was performed in final 50 μL of mixturecontaining PTase (20 μM PCPS, 5 nM hFVa, and 1 μM hFXa), 1.2 μM humanfactor 11 and varied concentration of the test compounds (5 μM to 0.021μM or lower concentration range). Reaction was started by addition ofPTase and incubated for 6 minutes at ambient temperature. Reaction wasstopped by addition of EDTA/buffer to final 10 mM. Activity of thrombin(product) was then measured in the presence of 0.1 mM of Spectrozyme-THas substrate at 405 nm for 5 minutes (10 seconds intervals) at ambienttemperature in a THEROmax microplate reader. Reactions were performed in96-well microtiter plates.

In the first step of the assay, 10 μL of diluted test compound (5×) orbuffer was added to the plates in duplicate. Then 10 μL of prothrombin(hFII) (5×) was added to each well. Next 30 μL PTase was added to eachwell, mix for about 30 seconds. The plates were then incubated atambient temperature for 6 minutes.

In the next step, 50 μL of 20 mM EDTA (in assay buffer) was added toeach well to stop the reaction. The resulting solutions were then mixedfor about 10 seconds. Then 100 μL of 0.2 mM spectrozyme was added toeach well. The thrombin reaction rate was then measured at 405 nm for 5minutes at 10 seconds intervals in a Molecular Devices microplatereader.

Calculations

Thrombin reaction rate was expressed as mOD/min. using OD readings fromthe five minute reaction. IC₅₀ values were calculated with the log-logitcurve fit program.

The compounds of the invention demonstrated the ability to inhibitpro-thrombinase when tested in this assay.

EXAMPLE 10 In Vivo Assay

The following assay demonstrates the ability of the compounds to act asanti-coagulants.

Male rats (250-330 g) were anesthetized with sodium pentobarbital (90mg/kg, i.p.) and prepared for surgery. The left carotid artery wascannulated for the measurement of blood pressure as well as for takingblood samples to monitor clotting variables (prothrombin time (PT) andactivated partial thromboplastin time (aPTT)). The tail vein wascannulated for the purpose of administering the test compounds (i.e.,the compounds of the invention and standards) and the thromboplastininfusion. The abdomen was opened via a mid-line incision and theabdominal vena cava was isolated for 2-3 cm distal to the renal vein.All venous branches in this 2-3 cm segment of the abdominal vena cavawere ligated. Following all surgery, the animals were allowed tostabilize prior to beginning the experiment. Test compounds wereadministered as an intravenous bolus (t=0). Three minutes later (t=3), a5-minute infusion of thromboplastin was begun. Two minutes into theinfusion (t=5), the abdominal vena cava was ligated at both the proximaland distal ends. The vessel was left in place for 60 minutes, afterwhich it was excised from the animal, slit open, the clot (if any)carefully removed, and weighed. Statistical analysis on the results wasperformed using a Wilcoxin-matched-pairs signed rank test.

The compounds of the invention, when tested in this assay, demonstratedthe ability to inhibit the clotting of the blood.

While the present invention has been described with reference to thespecific embodiments thereof, it should be understood by those skilledin the art that various changes may be made and equivalents may besubstituted without departing from the true spirit and scope of theinvention. In addition, many modifications may be made to adapt aparticular situation, material, composition of matter, process, processstep or steps, to the objective, spirit and scope of the presentinvention. All such modifications are intended to be within the scope ofthe claims appended hereto.

What is claimed is:
 1. A compound selected from the group consisting ofthe following formulae: ##STR12## wherein: Z¹ is --O--;Z² is --O--; R¹is --OR⁷ ; R² is --C(NH)NH₂ ; R³ is (1,2)-imidazolyl (optionallysubstituted by methyl) or (1,2)-imidazolinyl (optionally substituted bymethyl); R⁴ is hydrogen; R⁵ is hydrogen; and R⁶ is aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl); and R⁷ is hydrogen or alkyl; as a singlestereoisomer or a mixture thereof; or a pharmaceutically acceptable saltthereof.
 2. The compound of claim 1 wherein the compound is selectedfrom the formula (I): ##STR13##
 3. The compound of claim 2 wherein: R¹is hydroxy;R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).
 4. The compound of claim 3 wherein R⁶ is phenyl,i.e., namely the compound4-hydroxy-3-[[6-(3-(1-methylimidazolin-2-yl)phenoxy)-2-(phenyl)pyrimidin-4-yl]oxy]benzamidine.5. The compound of claim 1 wherein the compound is selected from theformula (II): ##STR14##
 6. The compound of claim 5 wherein: R¹ ishydroxy;R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).
 7. The compound of claim 6 wherein R⁶ is phenyl,i.e., namely, the compound4-hydroxy-3-[[4-(3-(1-methylimidazolin-2-yl)phenoxy)-6-(phenyl)pyrimidin-2-yl]oxy]benzamidine.8. The compound of claim 1 wherein the compound is selected from theformula (III): ##STR15##
 9. The compound of claim 8 wherein: R¹ ishydroxy;R³ is 1-methylimidazolin-2-yl; and R⁶ is aryl (optionallysubstituted by halo, haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy,aralkoxy, amino, dialkylamino, monoalkylamino, nitro, carboxy,alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl).
 10. The compound of claim 9 wherein R⁶ is phenyl,i.e., namely, the compound4-hydroxy-3-[[2-(3-(1-methylimidazolin-2-yl)phenoxy)-6-(phenyl)pyrimidin-4-yl]oxy]benzamidine.11. A pharmaceutical composition useful in treating a disease-state in ahuman,wherein the disease-state is characterized by thrombotic activityand is selected from the group consisting of thrombosis followingorthopedic surgery, coronary artery bypass graft and percutaneoustransluminal coronary angioplasty; thrombotic complications associatedwith acute promyelocytic leukemia, diabetes, and multiple myelomas;disseminated intravascular coagulation associated with septic shock;purpura fulminanas associated infection; adult respiratory distresssyndrome; unstable angina; and thrombotic complications associated withaortic valve or vascular prosthesis and transient ischemic attacks; andwherein the composition comprises a therapeutically effective amount ofa compound selected from the group consisting of the following formulae:##STR16## wherein: Z is --O--; Z² is --O--; R¹ is --OR⁷ ; R² is--C(NH)NH₂ ; R³ is (1,2)-imidazolyl (optionally substituted by methyl)or (1,2)-imidazolinyl (optionally substituted by methyl); R⁴ ishydrogen; R⁵ is hydrogen; R⁶ is aryl (optionally substituted by halo,haloalkyl, alkyl, aryl, aralkyl, hydroxy, alkoxy, aralkoxy, amino,dialkylamino, monoalkylamino, nitro, carboxy, alkoxycarbonyl,aminocarbonyl, monoalkylaminocarbonyl, or dialkylaminocarbonyl); and R⁷is hydrogen or alkyl; as a single stereoisomer or a mixture thereof; ora pharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable excipient.
 12. A method of treating a disease-state in ahuman,wherein the disease-state is characterized by thrombotic activityand is selected from the group consisting of thrombosis followingorthopedic surgery, coronary artery bypass graft and percutaneoustransluminal coronary angioplasty; thrombotic complications associatedwith acute promyelocytic leukemia, diabetes, and multiple myelomas;disseminated intravascular coagulation associated with septic shock;purpura fulminanas associated infection; adult respiratory distresssyndrome; unstable angina; and thrombotic complications associated withaortic valve or vascular prosthesis and transient ischemic attacks; andwherein the method comprises administering to the human in need thereofa therapeutically effective amount of a compound selected from the groupconsisting of the following formulae: ##STR17## wherein: Z¹ is --O--; Z²is --O--; R¹ is --OR⁷ ; R² is --C(NH)NH₂ ; R³ is (1,2)-imidazolyl(optionally substituted by methyl) or (1,2)-imidazolinyl (optionallysubstituted by methyl); R⁴ is hydrogen; R⁵ is hydrogen; R⁶ is aryl(optionally substituted by halo, haloalkyl, alkyl, aryl, aralkyl,hydroxy, alkoxy, aralkoxy, amino, dialkylamino, monoalkylamino, nitro,carboxy, alkoxycarbonyl, aminocarbonyl, monoalkylaminocarbonyl, ordialkylaminocarbonyl); and R⁷ is hydrogen or alkyl; as a singlestereoisomer or a mixture thereof; or a pharmaceutically acceptable saltthereof.